Recombinant Anti-Aly/Ref antibody [EPR17942] (ab202894)

Product name

Anti-Aly/Ref antibody [EPR17942]
See all Aly/Ref primary antibodies
Description

Rabbit monoclonal [EPR17942] to Aly/Ref
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: PC-3, HeLa, HepG2, HEK293 whole cell lysates; Human fetal brain, fetal heart and fetal kidney lysates; Mouse brain and heart lysates; rat brain and heart lysates. IHC-P: Human cervix carcinoma, Mouse cerebral cortex and rat liver tissues. ICC/IF: HeLa and C6 cells. Flow Cyt: HeLa cells. IP: HepG2 whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17942
Isotype

IgG
Research areas

Alternative Versions
- Anti-Aly/Ref antibody [EPR17942] - BSA and Azide free (ab240364)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab202894 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/150.
WB		1/2000. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).
IHC-P		1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/250.
IP		1/40.

Notes
Flow Cyt (Intra) 1/150.
WB 1/2000. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/250.
IP 1/40.

Function

Component of the THO subcomplex of the TREX complex. The TREX complex specifically associates with spliced mRNA and not with unspliced pre-mRNA. It is recruited to spliced mRNAs by a transcription-independent mechanism. Binds to mRNA upstream of the exon-junction complex (EJC) and is recruited in a splicing- and cap-dependent manner to a region near the 5' end of the mRNA where it functions in mRNA export. The recruitment occurs via an interaction between THOC4 and the cap-binding protein NCBP1. DDX39B functions as a bridge between THOC4 and the THO complex. TREX complex is essential for the export of Kaposi's sarcoma-associated herpesvirus (KSHV) intronless mRNAs and infectious virus production. The recruitment of the TREX complex to the intronless viral mRNA occurs via an interaction between KSHV ORF57 protein and THOC4. THOC4 in conjunction with THOC5 functions in NXF1-NXT1 mediated nuclear export of HSP70 mRNA.
Component of a splicing-dependent multiprotein exon junction complex (EJC) deposited at splice junction on mRNAs. The EJC is a dynamic structure consisting of a few core proteins and several more peripheral nuclear and cytoplasmic associated factors that join the complex only transiently either during EJC assembly or during subsequent mRNA metabolism. Acts as chaperone and promotes the dimerization of transcription factors containing basic leucine zipper (bZIP) domains and thereby promotes transcriptional activation. Plays a role in mRNA processing and export. May function as scaffold that mediates interactions between proteins and/or RNA. Is part of the exon junction complex that remains associated with spliced mRNA and plays an important role in mRNA export and nonsense-mediated RNA decay. Directs mRNA derived from Herpes simplex virus intron-less genes to the NXF1-mediated export pathway.
Sequence similarities

Belongs to the THOC4 family.
Contains 1 RRM (RNA recognition motif) domain.
Post-translational
modifications

Arg-50 and Arg-204 are dimethylated, probably to asymmetric dimethylarginine.
Phosphorylated upon DNA damage, probably by ATM or ATR.
Cellular localization

Nucleus. Nucleus speckle. Cytoplasm. Colocalizes with the core EJC, THOC4, NXF1 and DDX39B in the nucleus and nuclear speckles. Travels to the cytoplasm as part of the exon junction complex (EJC) bound to mRNA.
Target information above from: UniProt accession Q86V81 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 10189 Human
- Entrez Gene: 21681 Mouse
- Entrez Gene: 690585 Rat
- Omim: 604171 Human
- SwissProt: Q86V81 Human
- SwissProt: O08583 Mouse
- Unigene: 534385 Human
- Unigene: 1886 Mouse
Alternative names
- Ally of AML-1 and LEF-1 antibody
- Ally of AML1 and LEF1 antibody
- ALY antibody
- ALY/REF antibody
- Aly/REF export factor antibody
- BEF antibody
- bZIP enhancing factor antibody
- bZIP-enhancing factor BEF antibody
- REF antibody
- THO complex 4 antibody
- THO complex subunit 4 antibody
- Tho4 antibody
- thoc4 antibody
- THOC4_HUMAN antibody
- Transcriptional coactivator Aly/REF antibody
- Transcriptional coactivator antibody
see all

Western blot - Anti-Aly/Ref antibody [EPR17942] (ab202894)

All lanes : Anti-Aly/Ref antibody [EPR17942] (ab202894) at 1/10000 dilution

Lane 1 : PC-3 (Human prostate cancer cell line) whole cell lysate
Lane 2 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 4 : HEK293 (Human epithelial cells from embryonic kidney) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 27 kDa
Observed band size: 27 kDa

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Aly/Ref antibody [EPR17942] (ab202894)

All lanes : Anti-Aly/Ref antibody [EPR17942] (ab202894) at 1/2000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 27 kDa
Observed band size: 27 kDa

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Aly/Ref antibody [EPR17942] (ab202894)

All lanes : Anti-Aly/Ref antibody [EPR17942] (ab202894) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Rat brain lysate
Lane 4 : Rat heart lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

Predicted band size: 27 kDa
Observed band size: 27 kDa

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aly/Ref antibody [EPR17942] (ab202894)

Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling Aly/Ref with ab202894 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weakly cytoplasmic staining on Human cervix carcinoma tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aly/Ref antibody [EPR17942] (ab202894)

Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling Aly/Ref with ab202894 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weakly cytoplasmic staining on mouse cerebral cortex tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Aly/Ref antibody [EPR17942] (ab202894)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Aly/Ref with ab202894 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear and weakly cytoplasmic staining on rat liver tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Aly/Ref antibody [EPR17942] (ab202894)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Aly/Ref with ab202894 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows:
-ve control 1: ab202894 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Aly/Ref antibody [EPR17942] (ab202894)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cells) cells labeling Aly/Ref with ab202894 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weakly cytoplasmic staining on C6 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows:
-ve control 1: ab202894 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Flow Cytometry (Intracellular) - Anti-Aly/Ref antibody [EPR17942] (ab202894)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Aly/Ref with ab202894 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.
Immunoprecipitation - Anti-Aly/Ref antibody [EPR17942] (ab202894)

Aly/Ref was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma) whole cell lysate with ab202894 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab202894 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1: HepG2 whole cell lysate 10 µg (Input). Lane 2: ab202894 IP in HepG2 whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab202894 in HepG2 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 8 seconds.
Anti-Aly/Ref antibody [EPR17942] (ab202894)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

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Datasheet download

Download

Publishing research using ab202894? Please let us know so that we can cite the reference in this datasheet.

ab202894 has been referenced in 9 publications.

Hu L et al. Pharmacological inhibition of Ref-1 enhances the therapeutic sensitivity of papillary thyroid carcinoma to vemurafenib. Cell Death Dis 13:124 (2022). PubMed: 35136031
Wang L et al. SYVN1-MTR4-MAT2A Signaling Axis Regulates Methionine Metabolism in Glioma Cells. Front Cell Dev Biol 9:633259 (2021). PubMed: 33859984
Saleem I et al. The Mammalian Ecdysoneless Protein Interacts with RNA Helicase DDX39A To Regulate Nuclear mRNA Export. Mol Cell Biol 41:e0010321 (2021). PubMed: 33941617
He Y et al. Novel insights into the role of 5-Methylcytosine RNA methylation in human abdominal aortic aneurysm. Front Biosci (Landmark Ed) 26:1147-1165 (2021). PubMed: 34856760
Berson A et al. Drosophila Ref1/ALYREF regulates transcription and toxicity associated with ALS/FTD disease etiologies. Acta Neuropathol Commun 7:65 (2019). PubMed: 31036086
Park R & Miller G Epstein-Barr Virus-Induced Nodules on Viral Replication Compartments Contain RNA Processing Proteins and a Viral Long Noncoding RNA. J Virol 92:N/A (2018). PubMed: 30068640
Roundtree IA et al. YTHDC1 mediates nuclear export of N6-methyladenosine methylated mRNAs. Elife 6:N/A (2017). PubMed: 28984244
Boehringer A et al. ALS Associated Mutations in Matrin 3 Alter Protein-Protein Interactions and Impede mRNA Nuclear Export. Sci Rep 7:14529 (2017). PubMed: 29109432
Yang X et al. 5-methylcytosine promotes mRNA export - NSUN2 as the methyltransferase and ALYREF as an m5C reader. Cell Res 27:606-625 (2017). PubMed: 28418038

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (9)

Customer reviews and Q&As

Post-translational
modifications